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Protection for the skin

Extract from the Pariparoba exercises antioxidant action against the sun and should reach the market shortly

MIGUEL BOYAYANExtract ready for cosmetic useMIGUEL BOYAYAN

The Pariparoba, a shrub originating from the Atlantic Rain Forest, has shown in studies carried out at the Pharmaceutical Sciences College (FCF) of the University of São Paulo activities that protect against the ultraviolet rays of type UVB, the most damaging to the skin. The discovery resulted in a patent request and the interest of the company Natura, which disputed and won the bidding for the concession of the license for the use of this root extract in the development of products of cosmetic use (gel, cream, solar filters), with exclusivity for Brazil and abroad, carried out by the Invention Development Advise Group (Gadi), of the university.

“The potential antioxidant of the molecule found in the Pariparoba, responsible for the protection of the skin from free radicals, caught the attention of Natura”, says Jean Luc Gesztesi, a researcher in the company’s area of Research and Development (R&D). “For this reason we have the firm intention of using the extract in cosmetic products.” Meanwhile there is no fixed date for the launch, because the development of a product can be delayed from six months up to two years. However, the company already has a contract with a producer to provide the root extract of the Pariparoba (Pothomorphe umbellata) within certain specifications, such as the percentage of extract of the main active ingredient. Standardization is important in order to guarantee that the response to the vegetal extract is always the same, a characteristic named reproducibility.

“The Pariparoba has been used for a long time by popular medicine for the treatment of poor indigestion, liver illnesses such as jaundice, and burns”, says professor Silvia Berlanga de Moraes Barros, Cristina Dislich Ropke’s supervisor in her doctorate thesis, funded by FAPESP, which led to the discovery of the photo-protecting activity of the plant belonging to family of piperaceaes, mainly found in the States of São Paulo, Minas Gerais, Espírito Santo and the south of Bahia. In reality, research with the Pariparoba began with an investigation into the activity of hepatic protection attributed to the plant, which was not concluded. “A much closer look at the Pariparoba showed that it contained a substance that had previously been described by a phyto-chemistry group from the Chemistry Institute of USP in 1981”, professor says.

“The compound (4- nerolidylcatechol), a molecule found in the root extract of the plant, had some characteristics in its chemical structure very similar to that of alpha-tocopheral (vitamin E), an antioxidant used in cosmetic formulae that today are on the shelves for the prevention of skin aging, because it is an excellent protector of cellular membrane”, adds the researcher. The similarity of the characteristics led to an in vitro study to measure the antioxidant activity of the compound extracted from the root, carried out by professor Paulo Chanel Deodato de Freitas, also from FCF, at the time in which he was writing his doctorate degree, concluded in 1999. “As we verified that the compound had an activity much more potent than that of the alpha-tocopheral and had presented physical/chemical characteristics that could well justify the use in cosmetics, we decided to test it on the skin”, Silvia explains. The experiments were then done on hairless mice, a strain developed in animal breeding stations, which does not need to have their hairs removed, thus avoiding micro-lesions, on the skin and interfering in the research results.

The study had as its goal to show if the Pariparoba extract inhibited the spontaneous peroxidation of the skin – a chemical reaction also called oxidation -, that contributes to dermal aging and can occur as well through solar radiation. “In our model, we treated the animals with a formula that was very simple, without any technological treatment”, Silvia says. After the application and the excess having been removed, the product remained on the skin for an predetermined period. Next the researchers evaluated if the oxidation had been reduced more in the animals treated with the extract. “The microscope exams carried out on the mice showed that the compound prevented dermal aging”, says Silvia.

After this verification, the researchers decided to reproduce what happens in nature, with the exposure of the animals to ultraviolet radiation. At this stage a formula was chosen for the extract and a study on penetrating power carried out, which meant how much of the product stays on the skin and how much is absorbed. This information is important because part of the lesions caused by radiation, such as skin cancer, for example, concentrate themselves in the epidermis and part, such as photo-aging, in the layer of skin just below, or that is to say, the dermis. The study led to the formula in the form of a gel with the extract removed from the root. Though other parts of the plant also present the substance with the anti-oxidant, it is in the root that it is concentrated most intensely.

UVB rays
The next step was to expose the mice to UVB radiation, a part of the ultraviolet radiation that penetrates up to the epidermis. The UVB rays act on the surface of the earth between 10:00 AM and 3:00 PM and are the main culprits responsible for sun burns and, over time, for skin cancer. The treatment with the mice was done for ten minutes, four times per week, for a period of twenty two (22) weeks. “With this we saw that while the radiation triggers a massive increase in the cells of the epidermal layer, the so-called epithelial hyperplasia, that can lead to the development of carcinogenic cells, the animals treated with the extract presented reduced hyperplasia”, Silvia explains.

That was when the researcher realized that she was facing something new, because up until that moment the photo-protecting activity of the Pariparoba had not been proven in studies. As a consequence of this realization, during 2002 a patent request for the use of the extract of Pothomorphe umbellata in dermal cosmetic or pharmaceutical preparations for the prevention and combat of damage caused to the skin by excessive exposure to the sun’s ultraviolet rays and to artificial tanning lamps, as well as the alterations caused by chronological aging, was presented.

Other functions of the Pariparoba continue to be researched at the laboratory of the Pharmaceutical Sciences College of USP. The studies are centered on the bio-chemical mechanisms of the plant’s action. “The research is showing interesting results in the pharmaceutical area, but as yet they cannot be revealed”, Silvia points out. The link between the reduction of hyperplasia and the control of cellular growth could signify the bridge to medicines that regulate the development of carcinogenic cells.

In order to obtain the root extract that possessed proven laboratory results, the researchers made use of a classic technique of extraction at room temperature, called percolation. It is a totally a manual method, in which the roots, after being gathered, are dried and powdered. After they pass through a sieve, called a tamis, with controlled pore size, so that the extraction obeys a certain predetermined standard. Only then is the cold extraction done using ethanol, to remove the crude extract. By this method, the root has to remain constantly moistened by the ethanol so that the chemical substances are extracted by decantation. This is slow and takes between two to three days, until everything that needs to be extracted has been extracted. Only then is the removal of the molecule of interest done afterwards incorporated into the formula in the form of a gel or cream.

After the patent request had been complete, professor Silvia believed that the time had come for the university to open up a bidding process to choose a laboratory that could commercially produce cosmetic products based on the Pariparoba. The auction involved the payment of royalties on the products sales, which the company prefers not to reveal, and the transfer of technology on behalf of the university. USP and FAPESP, responsible for the concession of the doctoral scholarship, will each receive 50% of the contract with Natura. The two institutions are going to pass on part of this money to the researchers. The Foundation also financed the request for an international patent by way of the Support for Intellectual Property Program of the Patenting and Technology Licensing Center (Papi/Nuplitec).

The use of the Pariparoba extract is one of the projects with potential innovation that has its origin at the university and Natura betting on commercial utilization. For this reason, the company launched during the first semester of this year the program Natura Campus, which resulted in forty four (44) projects presented by the universities and research institutions to the company, of which thirteen (13) were selected and sent to FAPESP as part of the program Partnership for Technological Innovation (PITE). Of these, seven (7) were approved by the Foundation. For researcher Gesztesi, the generation of new knowledge at the university and applied by industry, could contribute to the development of differentiating products.

The Project
The use of the extract of Pothomorphe umbellata for the preparation of dermal cosmetic and/or pharmaceutical compounds (nº 03/08578-0); Modality Program of Intellectual Support (PAPI); Coordinator Silvia Berlanga de Moraes Barros; Investment R$ 6,400.00 (FAPESP)